RapidandAccurateSequencingofEnterovirusGenomesUsingMinIONNanoporeSequencer

摘要： ObjectiveKnowledgeofanenterovirusgenomesequenceisveryimportantinepidemiologicalinvestigationtoidentifytransmissionpatternsandascertaintheextentofanoutbreak.TheMinIONsequencerisincreasinglyusedtosequencevariousviralpathogensinmanyclinicalsituationsbecauseofitslongreads,portability,real-timeaccessibilityofsequenceddata,andverylowinitialcosts.However,informationislackingonMinIONsequencingofenterovirusgenomes.MethodsInthisproof-of-conceptstudyusingEnterovirus71(EV71)andCoxsackievirusA16(CA16)strainsasexamples,weestablishedanamplicon-basedwholegenomesequencingmethodusingMinION.Weexploredtheaccuracy,minimumsequencingtime,discriminationandhigh-throughputsequencingabilityofMinION,andcompareditsperformancewithSangersequencing.ResultsWithinthefirstminute(min)ofsequencing,theaccuracyofMinIONwas98.5%forthesingleEV71strainand94.12%-97.33%for10genetically-relatedCA16strains.Inaslittleas14min,99%identitywasreachedforthesingleEV71strain,andin17min(onaverage),99%identitywasachievedfor10CA16strainsinasinglerun.ConclusionMinIONissuitableforwholegenomesequencingofenteroviruseswithsufficientaccuracyandfinediscriminationandhasthepotentialasafast,reliableandconvenientmethodforroutineuse. （共9頁）